US4774467A - Method for recording nuclear magnetic resonance spectra - Google Patents

Method for recording nuclear magnetic resonance spectra Download PDF

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Publication number
US4774467A
US4774467A US06/937,949 US93794986A US4774467A US 4774467 A US4774467 A US 4774467A US 93794986 A US93794986 A US 93794986A US 4774467 A US4774467 A US 4774467A
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pulse
time reversal
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phase
spin
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Ole W. Sorensen
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Spectrospin AG
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Spectrospin AG
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01R—MEASURING ELECTRIC VARIABLES; MEASURING MAGNETIC VARIABLES
    • G01R33/00—Arrangements or instruments for measuring magnetic variables
    • G01R33/20—Arrangements or instruments for measuring magnetic variables involving magnetic resonance
    • G01R33/44—Arrangements or instruments for measuring magnetic variables involving magnetic resonance using nuclear magnetic resonance [NMR]
    • G01R33/46—NMR spectroscopy
    • G01R33/4633—Sequences for multi-dimensional NMR

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  • the present invention relates to a method for recording the nuclear magnetic resonance spectra of molecular spin systems between which there is an interactive effect in which the spin systems are excited by means of a 90° pulse and the resonance signals of the excited spin systems are not observed until the end of an evolution period t 1 that follows the 90° pulse.
  • signals S which are formed by totalling a plurality of different signals produced on the basis of the method according to the present invention are employed instead of the customary simple induction signals; these signals S are produced from the following equation, ##EQU7## in which s nq are individual induction signals that are obtained by varying n and q.
  • the method according to the present invention produces a chronological reversal of the evolution which occurs as a consequence of a scalar spin-spin coupling, with the result being homonuclear decoupling that does not possess the disadvantages of the constant-time experiment. Its principle is based upon the various dependencies of the various coherence transmission paths upon the flip angle ⁇ of a time reversal flip that is inserted in a suitable location.
  • the time reversal pulse which hereinafter will also be termed the TR pulse or TR element, produces a unique selection of those transmission paths which cause a chronological reversal of the evolution that was brought about by the J coupling.
  • the conditions for a chronological reversal can easily be indicated in the form of the spin multiplet components, expressed in the form of product operators. For example,
  • the method according to the present invention is of particular significance for homonuclear spin-spin decoupling (J decoupling) in conjunction with recording 2D spectrograms, it also enjoys general significance for chronological reversal of the evolutions caused by J couplings, and thus elimination of disturbing, J-caused signal attenuation.
  • the method according to the present invention can be employed in a versatile manner in conjunction with investigation of biological macromolecules by means of NMR spectroscopy, where conventional NMR spectra would otherwise display regions in which there is strong overlapping of the resonances. Elimination of multiplet splitting, i.e. refocussing of the J evolutions, can eliminate this overlapping and consequently open up new spectral regions for interpretation. Possibilities of application also exist in conjunction with heteronuclear spin systems and in 2D exchange spectroscopy in general.
  • the method can also be employed in conjunction with multiple quantum spectroscopy and in conjunction with indirect observation of magnetic resonance.
  • the time reversal (TR) pulse is comprised of two portions, i.e. of a first portion in the form of a 90° pulse having a phase of ⁇ n + ⁇ q and of a second portion, which follows the first portion, possibly at a brief chronological interval relative to the evolution period, in the form of a 90° pulse having a phase of ⁇ + ⁇ q .
  • ⁇ is the time that is required to alter the RF phase.
  • a typical value for ⁇ would be 5.10 -6 seconds.
  • the method according to the present invention is especially suitable for employment in conjunction with recording 2D spectra. Consequently, a preferred embodiment of the method according to the present invention calls for employment of pulse sequences that are suitable for recording two-dimensional nuclear magnetic resonance spectra in which the time reversal pulse is inserted and in which the phase of the RF oscillation of the excitation pulse that precedes the time reversal pulse is shifted by angle ⁇ n .
  • the pulse sequence that are suitable for recording two-dimensional nuclear magnetic resonance spectra in which the time reversal pulse is inserted and in which the phase of the RF oscillation of the excitation pulse that precedes the time reversal pulse is shifted by angle ⁇ n .
  • the invention is employed in conjunction with a method for investigating spin systems with heteronuclear coupling, in which a first class of nucleus is directly excited by a first pulse sequence and the excitation of the second class of nucleus that resulted through coupling is then queried by means of a second pulse sequence, the time reversal pulse is inserted into the first pulse sequence. It is obvious that here, too, it will again be necessary for the corresponding phase shift of the pulses preceding the TR pulse to occur and that the experiments can be repeated with the change in the various angles provided for by the present invention, and that the signals which are obtained and multiplied by the indicated weighting factor must be added together in order to obtain the signal that is free of the coupling.
  • Indirect investigation of spin systems with heteronuclear coupling can be performed in the known manner by exciting a first class of nucleus by means of a first pulse sequence and a second class of nucleus by means of a second pulse sequence in such a manner that a first 180° pulse following a first 90° pulse of the first pulse sequence is centered between two 90° pulses of the second pulse sequence.
  • the present invention can be employed in such a manner that the time reversal pulse is inserted in the first pulse sequence in a location that is chronologically in coincidence with the second 90° pulse of the second pulse sequence, in that the time reversal pulse is followed by a further 180° pulse at a chronological interval that is identical to the chronological interval between the first 90° pulse and the first 180° pulse, and in that scanning of the resonance signal provided by the first class of nucleus occurs at a chronological interval from the further 180° pulse that is identical to the chronological interval between the first 180° pulse and the time reversal pulse, so that the time reversal pulse is centered between the first 90° pulse and commencement of signal scanning.
  • FIG. 1 shows the diagram of the pulse sequence required for a 2D NOESY experiment, as modified according to the present invention
  • FIG. 2 shows a representation of a 2D NOE spectrum of deca-peptide-LHRH
  • FIGS. 3 and 4 show two regions of the spectrum illustrated in FIG. 2 on a larger scale, each both with and without reversing by means of a time reversal pulse;
  • FIGS. 5 to 10 show further diagrams of pulse sequences which have been modified according to the present invention.
  • FIG. 1 shows the diagram of a pulse sequence of the type employed in 2D NOE spectroscopy.
  • Typical of this pulse sequence are three 90° pulses 1, 2, 3, of which first 90° pulse 1 excites the spin system of the specimen to be examined, second 90° pulse 2 repeats the excitation following an evolution period t 1 in order to identify the influence of spin-spin couplings, while third 90° pulse 3 terminates a mixing period ⁇ m , which follows second 90° pulse 2. Following third 90° pulse 3, the resonance signal is received and recorded as an interferogram 4.
  • the RF oscillation of the first and last 90° pulses has phase positions ⁇ 1 and ⁇ 2 , respectively, relative to 90° pulse 2 which appears at the end of evolution period t 1 , while the phase-sensitive rectification of interferogram 4 with a phase ⁇ 1 + ⁇ 2 radio frequency signal alternately assumes the values of 0 and ⁇ .
  • a time reversal (TR) pulse is inserted between the first two 90° pulses, which define evolution period t 1 ; in the illustrated practical example, the TR pulse consists of two portions, i.e. of two 90° pulses 5 and 6, which are spaced at a chronological interval ⁇ one from the other, each being spaced t 1/2 from adjacent 90° pulses 1 and 2, respectively.
  • the RF oscillation of first 90° pulse 5 has a phase angle of ⁇ n + ⁇ q
  • the RF oscillation of second 90° pulse 6 has a phase position of ⁇ + ⁇ q .
  • Both 90° pulses 5 and 6 produce the effect of a pulse which causes the spin moments to twist about an angle ⁇ n and whose phase is shifted ⁇ q .
  • the phase angle of the first 90° pulse is shifted by ⁇ n , so that the RF oscillation of this pulse has a phase angle of ⁇ n + ⁇ 1 .
  • This phase cycle for ⁇ n is derived with consideration being given to the multiple quantum coherences (MQC) that are excited by 90° pulse 5 with phase ⁇ n + ⁇ q of the time reversal pulse. The multiple quantum coherences must be combined in the ratio of ##EQU9##
  • K represents the number of highest actually present MQC
  • p represents a current value between 0 and K.
  • B p results in the weight for the multiple quantum coherence of order p. If it is desired that J couplings or spins having K-1 or fewer J couplings be refocussed, phase ⁇ n must be increased in increments of ⁇ /N, where N>K.
  • the weightings for the individual increments of the ⁇ cycle are obtained through a Fourier transformation of values B p , i.e. ##EQU10##
  • n 0, 1, 2, . . . , N-1, N+1, . . . , 2N+1.
  • Phase ⁇ q must pass through the three values 0, 2/3 and 4/3, independently thereof.
  • the pair of pulses 5, 6 which form the time reversal pulse can thus also be viewed as being a multiple quantum filter.
  • the above-described phase cycle is then a combination of p-quantum-filtered spectra having weights of ##EQU12## for 0 ⁇ p ⁇ N equivalent.
  • NOESY-TR time reversal
  • zero quantum suppression which poses a known problem in the case of conventional NOE spectroscopy, is superfluous as a result of the fact that the J couplings have been refocussed.
  • TPPI time proportional phase increase
  • FIGS. 2 through 4 clearly illustrate the effect of the method according to the present invention when recording a 2D NOE spectrogram of deca-peptide-LHRH or p-Glu-His-Trp-Ser-Tyr-Gly 6 -Leu-Arg-Pro-Gly 10 -NH 2 .
  • the conventional NOESY diagram according to FIG. 2 displays heavy overlapping of the cross peaks in the two outlined regions 11, 12, which impedes allocation of these lines and preclude quantitative analysis of the NOE spectrogram.
  • regions 11 and 12 are again shown, on a larger scale.
  • the decoupling in the ⁇ 1 axis that was achieved through the application of the method according to the present invention produces the 2D spectrograms 13 and 14 which are shown in FIGS. 3b and 4b.
  • the illustrated spectrograms show the NH-C.sub. ⁇ H spectral region of a 300 MHz NOE spectrum. The recording was effected with 700 t 1 scans for a 2K ⁇ 2K data matrix packed with zeroes.
  • the major significance of the method according to the present invention results, among other things, from the fact that quantitative analysis of a NOESY spectrum, i.e. measurement of the signal amplitudes of the cross peaks, supplies direct information regarding atomic spacing. This information is of immense importance in understanding the tertiary structures of biological macromolecules. However quantitative analysis of this type is not possible if there is overlapping of the cross peaks in the conventional NOESY spectrum.
  • the NOESY-TR method according to the present invention represents what is thus far the only method through which the desired information can even be obtained in the case of otherwise overlapping cross peaks.
  • the method according to the present invention is suitable not only for reversing the evolution in the case of a J coupling in conjunction with 2D NOE spectroscopy, but can also be employed for other purposes.
  • the time reversal pulse with the associated phase shifts can be inserted into a plurality of different pulse sequences in which homonuclear or heteronuclear decoupling is desired.
  • FIGS. 5 through 10 show pulse sequences of the type that could be employed in other experiments.
  • FIG. 5 provides a general illustration of a pulse sequence for the observation of (homonuclear) exchanges in which, for observation of the exchanges, a 90° excitation pulse 21 is followed by an evolution period t 1 before the interferogram 22 is recorded after irradiation with a pair of pulses 24, which form a known double quantum filter having an independent phase cycle.
  • TR element 23 is inserted which, in turn, consists of two 90° portions having phase shifts of ⁇ n + ⁇ q and ⁇ + ⁇ q , in the above-described manner.
  • 90° pulse 21 that precedes TR element 23 has a phase shiit of ⁇ n .
  • mixing period ⁇ m is not terminated by a further 90° pulse at the end of evolution period t 1 , which, in turn, follows a 90° excitation pulse 31; instead, a 180° pulse 32 follows after one half of the mixing period ⁇ m /2, with 180° pulse 32 being followed by a further 90° pulse 33 at the end of mixing period ⁇ m , and with further 90° pulse 33 then being followed by recording of an interferogram 34.
  • a TR element 35 is also inserted after one half of the evolution period t 1 /2, as illustrated in FIG. 6.
  • 90° pulse 31 which precedes TR element 35, is, in turn, subjected to phase shift ⁇ n .
  • TOCSY totally correlated 2D spectrogram
  • FIG. 8 shows a diagram of the pulse sequence employed for recording a multiple quantum spectrogram.
  • evolution period t 1 is again initiated by a first 90° pulse 41 and terminated by a second 90° pulse 42.
  • a further 90° pulse 43 and a 180° pulse 44 are inserted prior to 90° pulse 41, which initiates evolution period t 1 .
  • An interferogram 45 is recorded following final 90° pulse 42.
  • a TR element 46 is again inserted into the pulse sequence after one half of the evolution period t 1 /2. In this case, it is necessary for not only 90° pulse 41, which initiates the evolution period, to be subjected to a phase shift of ⁇ n , but also the other pulses 43 and 44, which precede the TR element.
  • FIGS. 9 and 10 show pulse sequences of the type that are employed for investigating heteronuclear couplings and for indirect observation of magnetic resonance.
  • the H spins of the specimen to be examined are excited by two 90° pulses 51, 52, with an evolution period t 1 being initiated by 90° pulse 51 and terminated by 90° pulse 52.
  • a 90° pulse 53 is also irradiated for those X atoms of the spin system that are coupled with the H atoms.
  • These X atoms can consist of carbon -13, for example.
  • the interferogram 54 of the X atoms which is recorded after chronological interval ⁇ ' indicates the degree to which the X atoms have been excited through heteronuclear coupling by the previously excited H atoms during evolution period t 1 .
  • a TR element 55 is again inserted between the two 90° pulses 51, 52, which define evolution period t 1 , and the phase of first 90° pulse 51 is modified in the previously described manner.
  • Chronological interval ⁇ ' in which 180° pulses 56, 57 are centered, is employed, in the customary manner, for refocussing the antiphase signals prior to activation of decoupler 58.
  • both classes of atoms H and X are excited by one 90° pulse 61 or 62, respectively, one following the other, during a mixing period ⁇ m .
  • the chronologically subsequent 90° pulse which is employed for excitation of the X atoms is followed by a further 90° pulse 63 following evolution period t 1 , while the first class of atom H to be excited is irradiated with a 180° pulse 64, which follows the first 90° pulse after a period ⁇ m +t 1 /2.
  • the first class of atom H would normally be observed at time t 1 /2+ ⁇ m following 180° pulse 64.
  • a TR element 65 is inserted at time t 1 /2 following 180° pulse 64, with TR element 65 thus being inserted at the same time as 90° pulse 63, which terminates chronological interval t 1 and which is irradiated onto the other class of atom C.
  • This TR element 65 is followed at a chronological interval of ⁇ m +t 1 /2 by a further 180° pulse 66.
  • An interferogram 67 is recorded for first class of atom H at time t 1 /2 following irradiation of last 180° pulse 66.
  • TR element 65 is again centered in the total evolution period, as its chronological interval from 90° excitation pulse 61 amounts to ⁇ m +t 1 .
  • the two 90° and 180° pulses 61 and 64, which precede TR element 65 have an RF oscillation phase shift that amounts to a phase angle of ⁇ n .
  • the method according to the present invention can be implemented with all nuclear magnetic resonance spectrometers that have a generator for producing successive RF pulses with variable timing, as well as a variable phase position of the RF oscillation relative to a coherent reference oscillation, i.e. with all modern pulse Fourier transform spectrometers.

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  • Physics & Mathematics (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • High Energy & Nuclear Physics (AREA)
  • Condensed Matter Physics & Semiconductors (AREA)
  • General Physics & Mathematics (AREA)
  • Magnetic Resonance Imaging Apparatus (AREA)
  • Geophysics And Detection Of Objects (AREA)
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Cited By (13)

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WO1990006523A1 (de) * 1988-11-25 1990-06-14 Spectrospin Ag Verfahren zum selektiven anregen von nmr-signalen
WO1991009322A1 (de) * 1989-12-08 1991-06-27 Spectrospin Ag Hf-impuls-kaskade zur erzeugung von nmr-spektren
US5043664A (en) * 1988-11-03 1991-08-27 U.S. Philips Corp. Magnetic resonance spectroscopy method and device for performing the method
US5111819A (en) * 1988-11-25 1992-05-12 General Electric Nmr imaging of metabolites using a multiple quantum excitation sequence
US5229718A (en) * 1991-01-17 1993-07-20 Bruker Instruments, Inc. Method for increasing resolution in solid-state nmr spectra of abundant nuclei
US5668734A (en) * 1995-04-10 1997-09-16 The Uab Research Foundation Method for analyzing 2D transferred noesy spectra of molecules undergoing multistate conformational exchange
USD475799S1 (en) 2002-05-16 2003-06-10 Alert Lite Safety Products Co., Inc. Articulating arm light
US20100134104A1 (en) * 2006-07-31 2010-06-03 Schlumberger Technology Corporation Nuclear magnetic resonance measurement techniques in non-uniform fields
US20130021031A1 (en) * 2011-07-18 2013-01-24 Geoffrey Bodenhausen Fourier Tickling For Homonuclear Decoupling in NMR
US20140296695A1 (en) * 2007-11-07 2014-10-02 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Selective zero-quantum coherence transfer (sel-zqc) method for metabolite imaging in a poorly shimmed magnet field without susceptibility artifact
US20160154077A1 (en) * 2007-11-07 2016-06-02 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Msc-selmqc method for simultaneous mapping of polyunsaturated fatty acids, lactate and choline in high fat tissues
CN106872506A (zh) * 2017-03-15 2017-06-20 厦门大学 一种抵抗不均匀磁场的超快速核磁共振二维j谱方法
CN112824922A (zh) * 2019-11-21 2021-05-21 株式会社日立制作所 磁共振拍摄装置及其控制方法

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DE10144661C2 (de) * 2001-09-11 2003-08-14 Forschungsverbund Berlin Ev Vorrichtung und Verfahren zur Zuordnung der NMR-Signale von Polypeptiden
GB0308586D0 (en) * 2003-04-14 2003-05-21 Amersham Health R & D Ab Method and arrangements in NMR spectroscopy
CN109884107B (zh) * 2019-01-15 2020-07-31 厦门大学 一种测量同核间接偶合网络的方法
CN112834548B (zh) * 2021-01-08 2022-08-19 上海纽迈电子科技有限公司 一种交联密度测量方法及装置

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Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5043664A (en) * 1988-11-03 1991-08-27 U.S. Philips Corp. Magnetic resonance spectroscopy method and device for performing the method
WO1990006523A1 (de) * 1988-11-25 1990-06-14 Spectrospin Ag Verfahren zum selektiven anregen von nmr-signalen
US5111819A (en) * 1988-11-25 1992-05-12 General Electric Nmr imaging of metabolites using a multiple quantum excitation sequence
WO1991009322A1 (de) * 1989-12-08 1991-06-27 Spectrospin Ag Hf-impuls-kaskade zur erzeugung von nmr-spektren
US5285159A (en) * 1989-12-08 1994-02-08 Spectrospin Ag, Ind. RF pulse cascade for the generation of NMR spectra
US5229718A (en) * 1991-01-17 1993-07-20 Bruker Instruments, Inc. Method for increasing resolution in solid-state nmr spectra of abundant nuclei
US5668734A (en) * 1995-04-10 1997-09-16 The Uab Research Foundation Method for analyzing 2D transferred noesy spectra of molecules undergoing multistate conformational exchange
USD475799S1 (en) 2002-05-16 2003-06-10 Alert Lite Safety Products Co., Inc. Articulating arm light
US20100134104A1 (en) * 2006-07-31 2010-06-03 Schlumberger Technology Corporation Nuclear magnetic resonance measurement techniques in non-uniform fields
US7852077B2 (en) * 2006-07-31 2010-12-14 Schlumberger Technology Corporation Nuclear magnetic resonance measurement techniques in non-uniform fields
US9733326B2 (en) * 2007-11-07 2017-08-15 University of Pittsburgh—of the Commonwealth System of Higher Education Selective zero-quantum coherence transfer (Sel-ZQC) method for metabolite imaging in a poorly shimmed magnet field without susceptibility artifact
US20140296695A1 (en) * 2007-11-07 2014-10-02 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Selective zero-quantum coherence transfer (sel-zqc) method for metabolite imaging in a poorly shimmed magnet field without susceptibility artifact
US20160154077A1 (en) * 2007-11-07 2016-06-02 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Msc-selmqc method for simultaneous mapping of polyunsaturated fatty acids, lactate and choline in high fat tissues
US9915714B2 (en) * 2007-11-07 2018-03-13 University of Pittsburgh—of the Commonwealth System of Higher Education MSC-SELMQC method for simultaneous mapping of polyunsaturated fatty acids, lactate and choline in high fat tissues
US9086465B2 (en) * 2011-07-18 2015-07-21 Bruker Biospin Ag Fourier tickling for homonuclear decoupling in NMR
US20130021031A1 (en) * 2011-07-18 2013-01-24 Geoffrey Bodenhausen Fourier Tickling For Homonuclear Decoupling in NMR
CN106872506A (zh) * 2017-03-15 2017-06-20 厦门大学 一种抵抗不均匀磁场的超快速核磁共振二维j谱方法
CN106872506B (zh) * 2017-03-15 2018-05-18 厦门大学 一种抵抗不均匀磁场的超快速核磁共振二维j谱方法
CN112824922A (zh) * 2019-11-21 2021-05-21 株式会社日立制作所 磁共振拍摄装置及其控制方法
CN112824922B (zh) * 2019-11-21 2024-05-07 富士胶片医疗健康株式会社 磁共振拍摄装置及其控制方法

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DE3543123C2 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html) 1990-05-03
JPH0614016B2 (ja) 1994-02-23
DE3685305D1 (de) 1992-06-17
JPS62137554A (ja) 1987-06-20
EP0224854A2 (de) 1987-06-10
EP0224854B1 (de) 1992-05-13
EP0224854A3 (en) 1989-12-13

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